Hoby P. Hetherington

The flux from glucose to glutamate in the rat brain in vivo as determined by 1H-observed, 13C-edited NMR spectroscopy.

The rate of incorporation of carbon from [1-13C]glucose into the [4-CH2] and [3-CH2] of cerebral glutamate was measured in the rat brain in vivo by 1H-observed, 13C-edited (POCE) nuclear magnetic resonance (NMR) spectroscopy. Spectra were acquired every 98 s during a 60-min infusion of [1-13C]glucose. Complete time courses were obtained from six animals. The measured intensity of the unresolved [4-13CH2] resonances of glutamate and glutamine increased exponentially during the infusion and attained a steady state in ∼20 min with a first-order rate constant of 0.130 ± 0.010 min−1 (t1/2 = 5.3 ± 0.5 min). The appearance of the [3-13CH2] resonance in the POCE difference spectrum lagged behind that of the [4-13CH2] resonance and had not reached steady state at the end of the 60-min infusion (t1/2 = 26.6 ± 4.1 min). The increase observed in 13C-labeled glutamate represented isotopic enrichment and was not due to a change in the total glutamate concentration. The glucose infusion did not affect the levels of high-energy phosphates or intracellular pH as determined by 31P NMR spectroscopy. Since glucose carbon is incorporated into glutamate by rapid exchange with the tricarboxylic acid (TCA) cycle intermediate α-ketoglutarate, the rate of glutamate labeling provided an estimate of TCA cycle flux. We have determined the flux of carbon through the TCA cycle to be ≈1.4 μmol g−1 min−1. These experiments demonstrate the feasibility of measuring metabolic fluxes in vivo using 13C-labeled glucose and the technique of 1H-observed, 13C-decoupled NMR spectroscopy.

Effect of hypoglycemic encephalopathy upon amino acids, high-energy phosphates, and pHi in the rat brain in vivo: detection by sequential 1H and 31P NMR spectroscopy.

Abstract: Metabolic alterations in amino acids, high‐energy phosphates, and intracellular pH during and after insulin hypoglycemia in the rat brain was studied in vivo by 1H and 31P nuclear magnetic resonance (NMR) spectroscopy. Sequential accumulations of 1H and 31P spectra were obtained from a double‐tuned surface coil positioned over the exposed skull of a rat while the electroencephalogram was recorded continuously. The transition to EEG silence was accompanied by rapid declines in phosphocreatine, nucleoside triphosphate, and an increase in inorganic orthophosphate in 31P spectra. In 1H spectra acquired during the same time interval, the resonances of glutamate and glutamine decreased in intensity while a progressive increase in aspartate was observed. Following glucose administration, glutamate and aspartate returned to control levels (recovery half‐time, 8 min); recovery of glutamine was incomplete. An increase in lactate was detected in the 1H spectrum during recovery but it was not associated with any change in the intracellular pH as assessed in the corresponding 31P spectrum. Phosphocreatine returned to control levels following glucose administration, in contrast to nucleoside triphosphate and inorganic orthophosphate which recovered to only 80% and 200% of their control levels, respectively. These results show that the changes in cerebral amino acids and high‐energy phosphates detected by alternating the collection of 1H and 31P spectra allow for a detailed assessment of the metabolic response of the hypoglycemic brain in vivo.

Neurometabolism in human epilepsy

Purpose: Because of the large and continuous energetic requirements of brain function, neurometabolic dysfunction is a key pathophysiologic aspect of the epileptic brain. Additionally, neurometabolic dysfunction has many self‐propagating features that are typical of epileptogenic processes, that is, where each occurrence makes the likelihood of further mitochondrial and energetic injury more probable. Thus abnormal neurometabolism may be not only a chronic accompaniment of the epileptic brain, but also a direct contributor to epileptogenesis.

Effects of Acute Hyperammonemia on Cerebral Amino Acid Metabolism and pHi In Vivo, Measured by 1H and 31P Nuclear Magnetic Resonance

Abstract: The effects of an acute intravenous infusion of ammonium acetate on rat cerebral glutamate and glutamine concentrations, energy metabolism, and intracellular pH were measured in vivo with 1H and 31P nuclear magnetic resonance (NMR). The level of blood ammonia maintained by the infusion protocol used in this study (∼ 500 μM, arterial blood) did not cause significant changes in arterial Pco2, Po2, or pH. Cerebral glutamate levels fell to at least 80% of the preinfusion value, whereas glutamine concentrations increased 170% relative to the preinfusion controls. The fall in brain glutamate concentrations followed a time course similar to that of the rise of brain glutamine. There were no detectable changes in the content of phosphocreatine (PCr) or nucleoside triphosphates (NTP), within the brain regions contributing to the sensitive volume of the surface coil, during the ammonia infusion. Intracellular pH, estimated from the chemical shift of the inorganic phosphate resonance relative to the resonance of PCr in the 31P spectrum, was also unchanged during the period of hyperammonemia. 1H spectra, specifically edited to allow quantitation of the brain lactate content, indicated that lactate rose steadily during the ammonia infusion. Detectable increases in brain lactate levels were observed ∼ 10 min after the start of the ammonia infusion and by 50 min of infusion had more than doubled. Spectra acquired from rats that received a control infusion of sodium acetate were not different from the spectra acquired prior to the infusion of either ammonium or sodium acetate. The results reported here support earlier findings that an increased blood ammonia concentration has a pronounced effect on the brain concentrations of two important amino acids, glutamate and glutamine. They also provide in vivo evidence for the absence of a sustained alteration in either brain intracellular pH or in the concentration of high‐energy phosphate compounds during a period of acute hyperammonemia. The technique of in vivo NMR spectroscopy permits multiple, simultaneous measurements of important intermediary and energy metabolites in a single animal, in real time, prior to and during the systemic perturbation.

Role of very high order and degree B0 shimming for spectroscopic imaging of the human brain at 7 tesla

With the advent of ultrahigh field systems (7T), significant improvements in spectroscopic imaging (SI) studies of the human brain have been anticipated. These gains are dependent upon the achievable B0 homogeneity, both globally (σB  0Global , over the entire regions of interest or slice) and locally (σB  0Global , influencing the linewidth of individual SI voxels within the regions of interest). Typically the B0 homogeneity is adjusted using shim coils with spatial distributions modeled on spherical harmonics which can be characterized by a degree (radial dependence) and order (azimuthal symmetry). However, the role of very high order and degree shimming (e.g., 3rd and 4th degree) in MRSI studies has been controversial. Measurements of σB  0Global and σB  0Local were determined from B0 field maps of 64 × 64 resolution. In a 10 mm thick slice taken through the region of the subcortical nuclei, we find that in comparison to 1st–2nd degree shims, use of 1st–3rd and 1st–4th degree shims reduces σB  0Global by 29% and 55%, respectively. Using a SI voxel size of ∼1cc with an estimate of σB  0Local from 3 × 3 × 3 B0 map pixels in this subcortical region, the number of pixels with σB  0Local of less than 5 Hz increased from 24 to 59% with 1st–3rd and 1st–4th over 1st–2nd degree shims, respectively. Magn Reson Med, 2012.

RF shimming for spectroscopic localization in the human brain at 7 T.

Spectroscopic imaging of the human head at short echo times (≤15 ms) typically requires suppression of signals from extracerebral tissues. However, at 7 T, decreasing efficiency in B  1+ generation (hertz/watt) and increasing spectral bandwidth result in dramatic increases in power deposition and increased chemical shift registration artifacts for conventional gradient‐based in‐plane localization. In this work, we describe a novel method using radiofrequency shimming and an eight‐element transceiver array to generate a B  1+ field distribution that excites a ring about the periphery of the head and leaves central brain regions largely unaffected. We have used this novel B  1+ distribution to provide in‐plane outer volume suppression (>98% suppression of extracerebral lipids) without the use of gradients. This novel B  1+ distribution is used in conjunction with a double inversion recovery method to provide suppression of extracerebral resonances with T1s greater than 400 ms, while having negligible effect on metabolite ratios of cerebral resonances with T1s > 1000 ms. Despite the use of two adiabatic pulses, the high efficiency of the ring distribution allows radiofrequency power deposition to be limited to 3‐4 W for a pulse repetition time of 1.5 sec. The short echo time enabled the acquisition of images of the human brain, displaying glutamate, glutamine, macromolecules, and other major cerebral metabolites. Magn Reson Med, 2010.

Decreased hippocampal volume on MRI is associated with increased extracellular glutamate in epilepsy patients

Purpose: Temporal lobe epilepsy (TLE) is associated with smaller hippocampal volume and with elevated extracellular (EC) glutamate levels. We investigated the relationship between the hippocampal volume and glutamate in refractory TLE patients.

Hippocampal neurochemistry, neuromorphometry, and verbal memory in nondemented older adults

Background: Characterization of the behavioral correlates of neuromorphometry and neurochemistry in older adults has important implications for an improved understanding of the aging process. The objective of this study was to test the hypothesis that a measure of hippocampal neuronal metabolism was associated with verbal memory in nondemented older adults after controlling for hippocampal volume. Methods: 4-T MRI, proton magnetic resonance spectroscopy (1H MRS), and neuropsychological assessment were conducted in 48 older adults (23 women; mean age 81 years). Average hippocampal N-acetyl aspartate/creatine ratios (NAA/Cr) and hippocampal volumes were obtained. Neuropsychological evaluation included tests of verbal memory (Buschke and Grober Free and Cued Selective Reminding Test–Immediate Recall [FCSRT-IR], Wechsler Memory Scale–Revised Logical Memory subtest) and attention and executive function (Trail Making Test Parts A and B). Results: Linear regression analysis indicated that after adjusting for age, hippocampal NAA/Cr was a significant predictor of FCSRT-IR performance (β = 0.38, p = 0.01, R 2 = 0.21). Hippocampal volume was also a significant predictor of FCSRT-IR performance after adjusting for age and midsagittal area (β = 0.47, p = 0.01, R 2 = 0.24). In a combined model, hippocampal NAA/Cr (β = 0.33, p = 0.03) and volume (β = 0.35, p = 0.03) were independent predictors of FCSRT-IR performance, accounting for 30% of the variance in memory. Conclusions: These findings indicate that nondemented older adults with smaller hippocampal volumes and lower levels of hippocampal N-acetyl aspartate/creatine ratio metabolites perform more poorly on a test of verbal memory. The integrity of both the structure and metabolism of the hippocampus may underlie verbal memory function in nondemented elderly. GLOSSARY: BIMC = Blessed Information-Memory-Concentration test; CDR = Clinical Dementia Rating scale; EAS = Einstein Aging Study; FCSRT-IR = Buschke and Grober Free and Cued Selective Reminding Test–Immediate Recall; FOV = field of view; LM = Wechsler Memory Scale–Revised Logical Memory subtest; MCI = mild cognitive impairment; MR = magnetic resonance; MRS = magnetic resonance spectroscopy; NAA/Cr = N-acetyl aspartate/creatine ratio; TMTA = Trail Making Test Part A; TMTB = Trail Making Test Part B.

MRI- and MRS-Derived Hippocampal Correlates of Quantitative Locomotor Function in Older Adults

Gait measures have been shown to predict cognitive decline and dementia in older adults. Investigation of the neurobiology associated with locomotor function is needed to elucidate this relationship with cognitive abilities. This study aimed to examine magnetic resonance imaging (MRI; hippocampal volume)- and proton magnetic resonance spectroscopy (MRS; N-acetylaspartate to creatine (NAA/Cr) ratios)-derived hippocampal correlates of quantitative gait function (swing time (seconds), stride length (cm), and stride length variability (standard deviation)) in a subset of 48 nondemented older adults (24 males; mean age=81 years) drawn from the Einstein Aging Study, a community-based sample of individuals over the age of 70 residing in Bronx, New York. Linear regression analyses controlling for age were used to examine hippocampal volume and neurochemistry as predictors of gait function. We found that stride length was associated with hippocampal volume (beta=0.36, p=0.03; overall model R(2)=0.33, p=0.01), but not hippocampal neurochemistry (beta=0.09, p=0.48). Stride length variability was more strongly associated with hippocampal NAA/Cr (beta=-0.38, p=0.01; overall model R(2)=0.14, p=0.04) than hippocampal volume (beta=-0.33, p=0.08). Gait swing time was not significantly related to any neuroimaging measure. These relationships remained significant after accounting for memory and clinical gait impairments. These findings suggest that nondemented older adults exhibit increased stride length variability that is associated with lower levels of hippocampal neuronal metabolism, but not hippocampal volume. Conversely, decreased stride length is associated with smaller hippocampal volumes, but not hippocampal neurochemistry. Distinct neurobiological hippocampal substrates may support decreased stride length and increased stride length variability in older adults.

Short echo spectroscopic imaging of the human brain at 7T using transceiver arrays

Recent advances in magnet technology have enabled the construction of ultrahigh‐field magnets (7T and higher) that can accommodate the human head and body. Despite the intrinsic advantages of performing spectroscopic imaging at 7T, increased signal‐to‐noise ratio (SNR), and spectral resolution, few studies have been reported to date. This limitation is largely due to increased power deposition and B1 inhomogeneity. To overcome these limitations, we used an 8‐channel transceiver array with a short TE (15 ms) spectroscopic imaging sequence. Utilizing phase and amplitude mapping and optimization schemes, the 8‐element transceiver array provided both improved efficiency (17% less power for equivalent peak B1) and homogeneity (SD(B1) = ±10% versus ±22%) in comparison to a transverse electromagnetic (TEM) volume coil. To minimize the echo time to measure J‐modulating compounds such as glutamate, we developed a short TE sequence utilizing a single‐slice selective excitation pulse followed by a broadband semiselective refocusing pulse. Extracerebral lipid resonances were suppressed with an inversion recovery pulse and delay. The short TE sequence enabled visualization of a variety of resonances, including glutamate, in both a control subject and a patient with a Grade II oligodendroglioma. Magn Reson Med, 2009.